Harrisson F, van Hoof J, Vanroelen C
J Histochem Cytochem. 1986 Sep;34(9):1231-5. doi: 10.1177/34.9.3525669.
A solid-phase protease assay has been used to screen different commercial preparations of glycosaminoglycan-degrading enzymes for the presence of proteolytic activity. Proteases cannot be detected in preparations of testicular hyaluronidase and of chondroitinase at the concentration used for histochemical purposes. Commercial Streptomyces hyaluronidase contains proteolytic contaminants detectable at the concentration used for histochemistry. At higher concentrations, all preparations appear to be contaminated with proteases. The results obtained using this assay suggest that addition of a mixture of proteinase inhibitors containing N-ethylmaleimide, EDTA, pepstatin, and phenylmethanesulfonylfluoride or soybean trypsin inhibitor has little effect on the proteolytic activity of the glycosaminoglycan-degrading enzyme preparations, irrespective of the pH used. Moreover, the use of EDTA in this mixture is questionable. This study also describes two testicular hyaluronidase preparations that may be particularly useful in functional studies of the living organism, as they are only slightly contaminated.
已采用一种固相蛋白酶测定法，以筛查不同商业制剂中糖胺聚糖降解酶的蛋白水解活性。在所使用的用于组织化学目的的浓度下，在睾丸透明质酸酶制剂和软骨素酶制剂中未检测到蛋白酶。市售的链霉菌透明质酸酶含有在组织化学所用浓度下可检测到的蛋白水解污染物。在更高浓度下，所有制剂似乎都被蛋白酶污染。使用该测定法获得的结果表明，添加含有N - 乙基马来酰亚胺、乙二胺四乙酸（EDTA）、胃蛋白酶抑制剂和苯甲基磺酰氟或大豆胰蛋白酶抑制剂的蛋白酶抑制剂混合物，对糖胺聚糖降解酶制剂的蛋白水解活性影响很小，无论使用何种pH值。此外，在该混合物中使用EDTA存在疑问。本研究还描述了两种睾丸透明质酸酶制剂，因其污染轻微，可能在生物体的功能研究中特别有用。
